索拉非尼诱导人多发性骨髓瘤U266细胞凋亡及其机制研究

Research on Sorafenib Inducing Human Multiple Myeloma U266 Cell Apoptosis and Its mechanism

目的 :探讨索拉非尼对人多发性骨髓瘤细胞U266增殖和凋亡的影响,同时探索其作用机制。方法 :通过MTT法检测多发性骨髓瘤细胞U266在不同浓度的索拉非尼作用下的增殖抑制率;流式细胞术检测细胞凋亡率;同时设计引物VEGFR-2、VEGFR-3、PDGFR-β、c-Kit,采用Q-PCR检测索拉非尼处理后的多发性骨髓瘤细胞U266 m RNA表达水平,初步确定索拉非尼的作用靶点;最后用Western-blot检测PDGFR-β与VEGFR-3蛋白表达的情况。结果 :不同浓度的索拉非尼对多发性骨髓瘤细胞U266抑制增殖及促进凋亡作用各异,且抑制率和凋亡率均呈时间-浓度依赖性;索拉非尼作用48 h之后,与对照组比较VEGFR-2、VEGFR-3、PDGFR-β、c-Kit的m RNA表达均下调,其中VEGFR-3、PDGFR-β最为显著,Western-blot检测VEGFR-3、PDGFR-β蛋白表达量与Q-PCR结果一致。结论 :索拉非尼有抑制人多发性骨髓瘤细胞的增殖及促进其凋亡的作用,其机制可能与索拉非尼可以抑制细胞表面相关受体激活途径有关。

Objective This study aims to investigate the effects of sorafenib on multiple myeloma U266 cells proliferation and apoptosis, and to investigate its mechanism as references for excavating new molecular targeted drugs. Methods Using MTT test to evaluate the proliferation and inhibition rates of multiple myeloma U266 cells in variable concentrations of sorafenib. Using flow cytometry to assay apoptosis rate of tumor cells. Meanwhile, we designed primers VEGFR-2, VEGFR-3, PDGFR-β, c-Kit and used Q-PCR to assay m RNA expression level of multiple myeloma U266 cells after treatment of sorafenib to confirm the target of sorafenib preliminarily. At last, we adopted western blotting to detect the expression of PDGFR-β and VEGFR-3. Results It showed diversity on inhibition of proliferation and promotion of apoptosis of multiple myeloma U266 cells due to variable concentrations of sorafenib, and the inhibition rate and apoptosis rate were time-concentration dependent. Undergoing the treatments of Sorafenib after 48 h, compared with the control group, m RNA expression of VEGFR-2, VEGFR-3, PDGFR-β, c-Kit down-regulated, especially the VEGFR-3, PDGFR-β. Western blotting showed that expression of VEGFR-3 and PDGFR-β is consistent with Q-PCR results. Conclusion Sorafenib inhibits the proliferation of human multiple myeloma cells and promotes the apoptosis of the tumor cells, the mechanism of which may be that sorafenib can inhibit relevant cell surface receptor activation pathways.